中药药效成分群的合成生物学研究进展

doi:10.12211/2096-8280.2023-082

合成生物学 ›› 2024, Vol. 5 ›› Issue (3): 631-657.DOI: 10.12211/2096-8280.2023-082

中药药效成分群的合成生物学研究进展

查文龙, 卜兰, 訾佳辰

中国医学科学院&北京协和医学院药物研究所，天然药物活性物质与功能国家重点实验室，国家卫生健康委员会天然药物生物合成重点实验室，中国医学科学院酶与天然药物生物催化重点实验室，北京 100050

收稿日期:2023-11-21 修回日期:2024-03-18 出版日期:2024-06-30 发布日期:2024-07-12
通讯作者: 訾佳辰
作者简介:查文龙（1994—），男，博士后。研究方向为合成生物学，酶工程，代谢工程。E-mail： zhawenlong@imm.ac.cn
卜兰（1997—），女，博士研究生。研究方向为合成生物学，酶工程，代谢工程。E-mail：bulan@imm.ac.cn
訾佳辰（1978—），男，研究员，博士生导师。研究方向为合成生物学，酶工程，代谢工程。E-mail：zijiachen@imm.ac.cn
基金资助:
国家自然科学基金(82293682);中国博士后科学基金(2023M730332);中国医学科学院中央级公益性科研院所基本科研业务费(2021-RC350-009)

Advances in synthetic biology for producing potent pharmaceutical ingredients of traditional Chinese medicine

Wenlong ZHA, Lan BU, Jiachen ZI

State Key Laboratory of Bioactive Substance and Function of Natural Medicines，NHC Key Laboratory of Biosynthesis of Natural Products，CAMS Key Laboratory of Enzyme and Biocatalysis of Natural Drugs，Institute of Materia Medica，Chinese Academy of Medical Sciences & Peking Union Medical College，Beijing 100050，China

Received:2023-11-21 Revised:2024-03-18 Online:2024-06-30 Published:2024-07-12
Contact: Jiachen ZI

摘要/Abstract

摘要：

中药是中华民族的文化瑰宝，也是我国在新药创制领域的重要驱动力。许多中药材来源于稀缺物种，其药效物质的规模化获取困难，是制约中药新药创制研究的重要瓶颈。合成生物学的出现和快速发展为解决这一瓶颈问题提供了新的途径。目前，中药药效物质的合成生物学研究在单个药效分子的生物制备方法上取得了重要进展。中药的药效主要源于多成分作用的叠加和协同，所以药效成分群是中药药效物质的主要形式，然而针对药效成分群的合成生物学研究鲜有报道。建立中药药效成分群合成生物技术的关键是精确调控组成分子的比例，从而产出优质药效成分群。本文首先总结了挥发油、总皂苷、总黄酮、总木脂素、总生物碱等重要类型中药药效成分群形成机制的研究进展。然后，重点以檀香挥发油为例，介绍如何通过酶工程和代谢工程的联合运用实现药效成分群成分比例和产量的双重优化。最后，对中药药效成分群合成生物学领域的未来研究重点进行了展望，包括：（1）加强中药药效成分群生物合成途径解析方面的研究，重点深入阐明复杂药效成分群的形成机制；（2）加强代谢优化手段方面的创新研究，重点揭示未知代谢调控机制并基于此发展创新调控策略；（3）加强酶工程方法学的创新研究，重点发展新型理性设计和定向进化的联用技术以及人工智能辅助的酶工程技术。

关键词: 中药, 药效成分群, 合成生物学, 途径解析, 挥发油, 皂苷, 黄酮, 木脂素, 生物碱

Abstract:

Traditional Chinese medicine (TCM) is a treasure of Chinese civilization and also a good mine for drug development in China. Many TCM components come from rare biological species including plants, animals, and insects, making the preparation of these TCM pharmaceutical substances at large scales a bottleneck that substantially impedes TCM-based drug development. However, the rapid development of synthetic biology has provided a strategy for addressing this challenge. At present, significant progress has been made in the bio-production of individual TCM components, but the efficacy of TCM is mainly due to the synergistic effect of those ingredients, which are termed as pharmaceutical ingredient groups. Reports on constructing the bio-production platform of pharmaceutical ingredient groups are limited. Herein, we summarize research progress in the biogenic mechanism of important TCM pharmaceutical ingredient groups, such as volatile oils, saponins, flavonoids, lignans and alkaloids. Some individual components of pharmaceutical ingredient groups (e.g. ginsenosides) are synthesized by multiple branching pathways, which can be produced and formatted thereafter. On the other hand, some pharmaceutical ingredients such as sandalwood oil can be synthesized through single pathways/enzymatic reactions by engineering the key enzymes to optimize their ratio. We comment the strategy of combining enzyme engineering and metabolic engineering to optimize both the production of pharmaceutical ingredient groups and their ratio. At the end, we outline the prospect of synthetic biology research for producing pharmaceutical ingredient groups, including: (1) complete clarification of the biogenic mechanism of more complex pharmaceutical ingredient groups, (2) development of novel metabolic engineering approaches for breaking through homogenization of methodology, and (3) optimization of the catalytic characteristics of key synthetic enzymes by combining rational design and directed evolution.

Key words: traditional Chinese medicine, pharmaceutical ingredient groups, synthetic biology, pathway elucidation, volatile oil, saponins, flavonoids, lignans, alkaloids

中图分类号:

R932

查文龙, 卜兰, 訾佳辰. 中药药效成分群的合成生物学研究进展[J]. 合成生物学, 2024, 5(3): 631-657.

Wenlong ZHA, Lan BU, Jiachen ZI. Advances in synthetic biology for producing potent pharmaceutical ingredients of traditional Chinese medicine[J]. Synthetic Biology Journal, 2024, 5(3): 631-657.

图/表 12

图1 檀香挥发油成分群的生物合成途径（DXS—1-脱氧-D-木酮糖-5-磷酸合酶；DXR—1-脱氧-D-木酮糖-5-磷酸还原异构酶；MCT—2-C-甲基-D-赤藻醇-4-磷酸胞苷酰转移酶；CMK—4-（5′-焦磷酸胞苷）-2-C-甲基-D-赤藓醇激酶；MCS—2-C-甲基-D-赤藓醇-2，4-环焦磷酸合成酶；HDS—1-羟基-2-甲基-2-丁烯-4-焦磷酸合成酶；HDR—1-羟基-2-甲基-2-丁烯-4-焦磷酸还原酶；IDI—异戊烯基焦磷酸异构酶；AACT—乙酰辅酶A酰基转移酶；HMGS—3-羟基-3-甲基戊二酰辅酶A合成酶；HMGR—3-羟基-3-甲基戊二酰辅酶A还原酶；MK—甲羟戊酸激酶；PMK—磷酸甲羟戊酸激酶；MPD—焦磷酸甲羟戊酸脱羧酶）

Fig. 1 Biosynthetic pathway for the production of sandalwood oil(DXS—1-deoxy-D-xylulose 5-phosphate synthase; DXR—1-deoxy-D-lxylulose 5-phosphate reductoisomerase; MCT—2-C-methyl-D-erythritol-4-phosphate cytidyltransferase; CMK—4-(cytidine 5′-diphospho)-2-C-methyl-D-erythritol kinase; MCS—2-C-methyl-D-erythritol 2,4-diphosphate synthase; HDS—1-hydroxy-2-methyl-2-E-butenyl-4-diphosphate synthase; HDR—1-hydroxy-2-methyl-2-butenyl-4-diphosphate reductase; IDI—isopentenyl pyrophosphate isomerase; AACT—acetyl-coenzyme A acetyltransferase; HMGS—3-hydroxy-3-methylglutaryl-CoA synthase; HMGR—3-hydroxy-3-methylglutaryl-CoA reductase; MK—mevalonate kinase; PMK—phosphate mevalonate kinase; MPD—mevalonate pyrophosphate decarboxylase)

表1 檀香醇生物合成相关细胞色素P450酶

Table 1 Cytochrome P450 enzymes for santalols biosynthesis

基因	底物	产物	参考文献
CYP76F41、CYP76F42、CYP76F39v1	α-檀香烯、 β-檀香烯、 epi-β-檀香烯、 exo-α-香柠檬烯	Z/E-α-檀香醇、 Z/E-β-檀香醇、 Z/E-epi-β-檀香醇、 Z/E-exo-α-香柠檬醇	[42]
CYP76F39v2		Z/E-α-檀香醇、 Z/E-β-檀香醇、 E-epi-β-檀香醇、 Z/E-exo-α-香柠檬醇
CYP76F40		Z-α-檀香醇、 E-β-檀香醇、 E-exo-α-香柠檬醇
CYP76F37v1、CYP76F37v2、CYP76F38v1、CYP76F38v2		E-α-檀香醇、 E-β-檀香醇、 E-exo-α-香柠檬醇
CYP736A167		Z-α-檀香醇、 Z-β-檀香醇、 Z-epi-β-檀香醇、 Z-exo-α-香柠檬醇	[43]

图2 广藿香挥发油成分群的生物合成途径

Fig. 2 Biosynthetic pathways for patchouli oils

图3 人参皂苷类成分群的生物合成途径

Fig. 3 Biosynthetic pathway for ginsenosides

图4 黄芪皂苷苷元合成和糖基化途径

Fig. 4 Synthesis and glycosylation of astragaloside aglucones in Astragali Radix

图5 黄芪皂苷类成分的乙酰化修饰

Fig. 5 Acetylation of astragalosides in Astragali Radix

图6 黄芩黄酮类成分群的生物合成途径

Fig. 6 Biosynthetic pathway for flavonoids in S. baicalensis

图7 淫羊藿黄酮类成分群的生物合成途径

Fig. 7 Biosynthetic pathway for flavonoids in Epimedii Folium

图8 板蓝根木脂素类成分群的生物合成途径

Fig. 8 Biosynthetic pathway for lignans in I. indigotica

图9 颠茄托品烷生物碱类成分群的生物合成途径

Fig. 9 Biosynthetic pathway for tropane alkaloids in A. belladonna

图10 檀香挥发油的合成生物学方法

Fig. 10 Production of sandalwood oil through synthetic biology approach

图11 水飞蓟宾类成分群的合成生物学方法

Fig. 11 Production of silybin constituent groups through synthetic biology approach

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中药药效成分群的合成生物学研究进展

Advances in synthetic biology for producing potent pharmaceutical ingredients of traditional Chinese medicine

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